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1010 electron microscope

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The 1010 electron microscope is a high-performance imaging device designed for detailed analysis of microscopic samples. It utilizes a focused beam of electrons to generate magnified images, allowing for the examination of fine structures and features at the nanoscale level. The core function of the 1010 electron microscope is to provide researchers and scientists with a powerful tool for advanced material characterization, structural analysis, and imaging of a wide range of specimens.

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Lab products found in correlation

Embed 812, by Electron Microscopy Sciences (9 mentions) Advantage image capture software, by Hamamatsu Photonics (3 mentions) Bx53 microscope, by Hamamatsu Photonics (1 mentions) Orca r2 c10600 digital camera, by Hamamatsu Photonics (1 mentions) Amt advantage image capture software, by Hamamatsu Photonics (1 mentions) Digital camera, by Hamamatsu Photonics (1 mentions) Pam3csk4, by InvivoGen (1 mentions) Durcupan acm resin, by Merck Group (1 mentions) Ultracut uct, by Leica (1 mentions)

12 protocols using 1010 electron microscope

1

Ultrastructural Analysis of Sciatic Nerve

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Mice were perfused intravascularly with 2.5% glutaraldehyde and 4% paraformaldehyde in 0.1M cacodylate buffer pH 7.4. Sciatic nerves and nerve roots were removed and incubated in the same buffer for 90 min at room temperature then overnight at 4°C, with rotation. Specimens were post-fixed for 1 hr with OsO4, dehydrated then embedded in araldite. Semi-thin (0.5 μm) and ultrathin (70 nm) section were cut using an ultramicrotome, and stained with toluidine blue or uranyl acetate and lead citrate, respectively. Semi-thin sections were examined using bright-field on an Olympus BX53 microscope, and images captured using Hamamatsu ORCA R2 C10600 digital camera and Metamorph software.
For analysis of the number of myelin profiles in control vs mutant sciatic nerves, semi-thin sections from proximal-, mid- and distal regions of the sciatic nerve were taken, and two non- overlapping images of each section were taken using the 60x objective: these images were termed ‘field of view’ (FOV). The average number of myelin profiles in each FOV was calculated, using ImageJ software. For electron microscopy, stained ultrathin sections were examined using a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Grove M., Kim H., Santerre M., Krupka A.J., Han S.B., Zhai J., Cho J.Y., Park R., Harris M., Kim S., Sawaya B.E., Kang S.H., Barbe M.F., Cho S.H., Lemay M.A, & Son Y.J. (2017). YAP/TAZ initiate and maintain Schwann cell myelination. eLife, 6, e20982.
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2

Ultrastructural Analysis of Carotid Arteries

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Carotid arteries for TEM were fixed overnight at 4°C in 0.1M sodium cacodylate buffer, pH 7.4 containing 2.5% glutaraldehyde and 2.0% paraformaldehyde. After subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide for 1 h at room temperature and rinsed in water before en bloc staining with 2% uranyl acetate. Briefly, after dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences). Thin cross sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software. Images of artery cross sections were taken at increasing magnification; collagen abundance was evaluated as described in Supplemental Data 1.
von Kleeck R., Roberts E., Castagnino P., Bruun K., Brankovic S.A., Hawthorne E.A., Xu T., Tobias J.W, & Assoian R.K. (2021). Arterial stiffness and cardiac dysfunction in Hutchinson–Gilford Progeria Syndrome corrected by inhibition of lysyl oxidase. Life Science Alliance, 4(5), e202000997.
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3

Electron Microscopic Tissue Preparation

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Tissues for electron microscopic examination were fixed with 2.5% glutaraldehyde, 2% paraformaldehyde in 0.1 M sodium cacodylate buffer, pH 7.4, overnight at 4 oC. After subsequent buffer washes, the samples were post -fixed in 2% osmium tetroxide for 1 hour at room temperature, and rinsed in deionized water prior to en bloc staining with 2% uranyl acetate. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Rybakovsky E., Buleza N.B., Hoxha K., DiGuilio K.M., McCluskey E.S., Friday C.L., Callaghan P.J., Moskalenko D.V., Zuo B., Thomas S, & Mullin J.M. (2018). Spontaneous and cytokine-induced hole formation in epithelial cell layers: Implications for barrier function studies with the gingival cell culture, Gie-3B11, and other epithelial models. Trends in cell & molecular biology, 13, 99-114.
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4

Electron Microscopic Analysis of Cardiomyocytes

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Cardiomyocytes for electron microscopic examination were sorted directly into fixative containing 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1M sodium cacodylate buffer, pH 7.4, overnight at 4°C. After subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide for 1 hour at room temperature, and rinsed in DH2O prior to en bloc staining with 2% uranyl acetate. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Windmueller R., Leach J.P., Babu A., Zhou S., Morley M.P., Wakabayashi A., Petrenko N.B., Viatour P, & Morrisey E.E. (2020). Direct Comparison of Mononucleated and Binucleated Cardiomyocytes Reveals Molecular Mechanisms Underlying Distinct Proliferative Competencies. Cell reports, 30(9), 3105-3116.e4.
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5

Ultrastructural Analysis of Fibroblasts

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For these studies, we utilized the non-disease control 1 and 2, VLCAD mutant 1 and 3, and TFP mutant 1 and 3 fibroblasts. The cells were grown in T75 flasks at 90% confluence (about 1 × 106 cells). Cells were harvested by trypsinization, fixed with 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1 M sodium cacodylate buffer, pH 7.4, overnight at 4 °C. After several washes, the samples were fixed with a 2.0% osmium tetroxide for 1 h at room temperature. Samples were rinsed with ddH2O and treated with 2% uranyl acetate. After dehydration through a graded ethanol series, the cells were embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Raimo S., Zura-Miller G., Fezelinia H., Spruce L.A., Zakopoulos I., Mohsen A.W., Vockley J, & Ischiropoulos H. (2021). Mitochondrial morphology, bioenergetics and proteomic responses in fatty acid oxidation disorders. Redox Biology, 41, 101923.
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6

Ultrastructural Analysis of Autophagic Vesicles

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For TEM, cells and tissues were fixed and analyzed as described previously [29 (link)]. Briefly, tissues were fixed in 2.5% (w/v) glutaraldehyde and 2.0% paraformaldehyde in 0.1 M cacodylate buffer (pH 7.4) at 4°C overnight. After buffer wash, the samples were post-fixed in 2% osmium tetroxide for 1 h, washed again in buffer, and dehydrated in a graded ethanol series. Samples were treated with several changes of hexamethyldisilazane (HMDS) and then allowed to air dry prior to mounting and sputter-coating with gold. TEM examinations were made, and TEM photos were taken with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage imaging software. AVs were identified based on published ultrastructural features [30 (link)], most significantly the presence of double or multiple membranes as well as vesicles which contain cytoplasmic material. Counts were performed by two independent investigators and averaged for each cell. Cells counts were then averaged and statistical testing performed as described.
Kong J., Whelan K.A., Laczkó D., Dang B., Monroig A.C., Soroush A., Falcone J., Amaravadi R.K., Rustgi A.K., Ginsberg G.G., Falk G.W., Nakagawa H, & Lynch J.P. (2015). Autophagy levels are elevated in Barrett’s esophagus and promote cell survival from acid and oxidative stress. Molecular carcinogenesis, 55(11), 1526-1541.
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7

Transmission Electron Microscopy Protocol

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Tissues and cells for electron microscopic examination were fixed with 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1 mol/L sodium cacodylate buffer, pH 7.4, overnight at 4°C. After subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide with 1.5% K3Fe(CN)6 for 1 hour at room temperature, and rinsed in DH2O prior to en bloc staining with 2% uranyl acetate. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Dierickx P., Zhu K., Carpenter B.J., Jiang C., Vermunt M.W., Xiao Y., Luongo T.S., Yamamoto T., Martí-Pàmies Í., Mia S., Latimer M., Diwan A., Zhao J., Hauck A.K., Krusen B., Nguyen H.C., Blobel G.A., Kelly D.P., Pei L., Baur J.A., Young M.E, & Lazar M.A. (2021). Circadian REV-ERBs repress E4bp4 to activate NAMPT-dependent NAD+ biosynthesis and sustain cardiac function. Nature cardiovascular research, 1(1), 45-58.
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8

Electron Microscopy Analysis of Salmonella-Infected Macrophages

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Two days prior to experimentation, 2 × 106 cells THP-1 cells were replated in 10-cm dishes in media without antibiotics and incubated with phorbol 12-myristate 13-acetate (PMA) for 24 hours to allow differentiation into macrophages. Then, macrophages were primed with 100 ng/mL Pam3CSK4 (Invivogen) for 16 hours prior to bacterial infection. Cells were infected with WT Salmonella at an MOI of 20 as described above. At 8 hpi, the media was aspirated, and the cells were fixed with 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1M sodium cacodylate buffer, pH 7.4. Then, at the Electron Microscopy Resource Laboratory in the Perelman School of Medicine, after subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide with 1.5% K3Fe(CN)6 for 1 hour at room temperature, and rinsed in dH2O. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and SATO lead and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage NanoSprint500 software.
Egan M.S., O’Rourke E.A., Mageswaran S.K., Zuo B., Martynyuk I., Demissie T., Hunter E.N., Bass A.R., Chang Y.W., Brodsky I.E, & Shin S. (2023). Inflammasomes primarily restrict cytosolic Salmonella replication within human macrophages. bioRxiv.
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9

Transmission Electron Microscopy Protocol

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Tissues and cells for electron microscopic examination were fixed with 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1 mol/L sodium cacodylate buffer, pH 7.4, overnight at 4°C. After subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide with 1.5% K3Fe(CN)6 for 1 hour at room temperature, and rinsed in DH2O prior to en bloc staining with 2% uranyl acetate. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Dierickx P., Zhu K., Carpenter B.J., Jiang C., Vermunt M.W., Xiao Y., Luongo T.S., Yamamoto T., Martí-Pàmies Í., Mia S., Latimer M., Diwan A., Zhao J., Hauck A.K., Krusen B., Nguyen H.C., Blobel G.A., Kelly D.P., Pei L., Baur J.A., Young M.E, & Lazar M.A. (2021). Circadian REV-ERBs repress E4bp4 to activate NAMPT-dependent NAD+ biosynthesis and sustain cardiac function. Nature cardiovascular research, 1(1), 45-58.
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10

Electron Microscopic Analysis of Cardiomyocytes

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Cardiomyocytes for electron microscopic examination were sorted directly into fixative containing 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1M sodium cacodylate buffer, pH 7.4, overnight at 4°C. After subsequent buffer washes, the samples were post-fixed in 2.0% osmium tetroxide for 1 hour at room temperature, and rinsed in DH2O prior to en bloc staining with 2% uranyl acetate. After dehydration through a graded ethanol series, the tissue was infiltrated and embedded in EMbed-812 (Electron Microscopy Sciences, Fort Washington, PA). Thin sections were stained with uranyl acetate and lead citrate and examined with a JEOL 1010 electron microscope fitted with a Hamamatsu digital camera and AMT Advantage image capture software.
Windmueller R., Leach J.P., Babu A., Zhou S., Morley M.P., Wakabayashi A., Petrenko N.B., Viatour P, & Morrisey E.E. (2020). Direct Comparison of Mononucleated and Binucleated Cardiomyocytes Reveals Molecular Mechanisms Underlying Distinct Proliferative Competencies. Cell reports, 30(9), 3105-3116.e4.
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